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dc.contributorVirginia Tech. School of Biomedical Engineering and Sciencesen_US
dc.contributorVirginia Tech. Department of Biological Sciencesen_US
dc.contributorVirginia Tech. Department of Chemical Engineeringen_US
dc.contributor.authorCao, Zhenningen_US
dc.contributor.authorGeng, Shuoen_US
dc.contributor.authorLi, Liwuen_US
dc.contributor.authorLu, Changen_US
dc.identifier.citationCao, Z., Geng, S., Li, L., & Lu, C. (2014). Detecting intracellular translocation of native proteins quantitatively at the single cell level. Chemical Science, 5(6), 2530-2535. doi: 10.1039/C4SC00578C
dc.description.abstractThe intracellular localization and movement (i.e. translocation) of proteins are critically correlated with the functions and activation states of these proteins. Simple and accessible detection methods that can rapidly screen a large cell population with single cell resolution have been seriously lacking. In this report, we demonstrate a simple protocol for detecting translocation of native proteins using a common flow cytometer which detects fluorescence intensity without imaging. We sequentially conducted chemical release of cytosolic proteins and fluorescence immunostaining of a targeted protein. The detected fluorescence intensity of cells was shown to be quantitatively correlated to the cytosolic/nuclear localization of the protein. We used our approach to detect the translocation of native NF-_B (an important transcription factor) at its native expression level and examine the temporal dynamics in the process. The incorporation of fluorescence immunostaining makes our approach compatible with the analysis of cell samples from lab animals and patients. Our method will dramatically lower the technological hurdle for studying subcellular localization of proteins.
dc.publisherThe Royal Society of Chemistry
dc.rightsCreative Commons Attribution-NonCommercial 3.0 Unported
dc.subjectFluorescence microscopyen_US
dc.subjectFlow cytometry analysisen_US
dc.subjectStandard fluorescence immunostainingen_US
dc.subjectWestern blottingen_US
dc.titleDetecting intracellular translocation of native proteins quantitatively at the single cell levelen_US
dc.typeArticle - Refereeden_US
dc.description.notesSupplementary information is included in a separate file
dc.title.serialChemical Science

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Creative Commons Attribution-NonCommercial 3.0 Unported
License: Creative Commons Attribution-NonCommercial 3.0 Unported